| 引用本文:龚竹韵1，杨晓丹1，程序2，孙怀艳2，高翔1.葛根芩连汤调控微RNA-542-3p对溃疡性结肠炎的保护机制[J].世界中医药,2022,(16):. |
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| 葛根芩连汤调控微RNA-542-3p对溃疡性结肠炎的保护机制 |
| Protective Mechanism of Gegen Qinlian Decoction against Ulcerative Colitis by Regulating MicroRNA-542-3p |
| 投稿时间：2022-03-23 |
| DOI：10.3969/j.issn.1673-7202.2022.16.016 |
| 中文关键词: 葛根芩连汤 微RNA-542-3p 溃疡性结肠炎 炎症介质 氧化应激 美沙拉嗪 酶联免疫检测 实时荧光定量PCR |
| English Keywords:Gegen Qinlian Decoction miR-542-3p Ulcerative colitis Inflammatory factors Oxidative stress Mesalazine Enzyme-linked immunosorbent assay Real-time fluorescence quantitative PCR |
| 基金项目:全国中医临床特色技术传承骨干人才培训项目(国中医药人教函[2019]36号) |
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| 中文摘要: |
| 目的:探讨葛根芩连汤上调微RNA-542-3p(miR-542-3p)表达保护溃疡性结肠炎(UC)大鼠的机制研究。方法:选取8~10周龄SD大鼠经UC造模及鉴定后分成模型组、美沙拉嗪组、葛根芩连汤低剂量、中剂量、高剂量组，同批次等体质量健康大鼠作为空白对照组，每组16只。UC结肠细胞进入对数生长期后消化传代至24孔板培养，每孔加入2×105个细胞，用Lipofectamine 3000转染miR-NC、miR-542-3p、anti-miR-NC、anti-miR-542-3p质粒至UC大鼠结肠上皮细胞并分组，miR-NC、miR-542-3p组细胞经含10%生理盐水处理24 h，而anti-miR-NC、anti-miR-542-3p组用含10%葛根芩连汤处理24 h，处理结束后分析细胞上清液炎症介质和氧化应激相关指标的变化。酶联免疫吸附试验法(ELISA)检测大鼠结肠组织和细胞肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)含量;化学比色法检测丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)含量;实时荧光定量聚合酶链式反应(RT-qPCR)检测miR-542-3p表达水平。结果:与模型组比较,葛根芩连汤中剂量、高剂量组TNF-α、IL-6、IL-8含量显著降低(P<0.05)。与模型组比较,葛根芩连汤中剂量、高剂量组MDA水平显著降低,SOD、GSH-PX水平显著升高(P<0.05)。与模型组比较,葛根芩连汤中、高剂量组miR-542-3p表达水平显著升高(P<0.05)。与UC+miR-NC组比较,UC+miR-542-3p组细胞miR-542-3p表达水平显著升高;TNF-α、IL-6、IL-8含量显著降低;MDA水平显著降低以及SOD、GSH-Px水平显著升高(P<0.05)。与UC+anti-miR-NC+葛根芩连汤组比较,UC+anti-miR-542-3p+葛根芩连汤组细胞miR-542-3p表达水平显著降低;TNF-α、IL-6、IL-8含量显著升高;MDA水平显著升高以及SOD、GSH-Px水平显著降低(P<0.05)。结论:葛根芩连汤上调miR-542-3p表达保护UC大鼠,消除肠道炎症。 |
| English Summary: |
| To investigate the mechanism of Gegen Qinlian Decoction in protecting ulcerative colitis(UC) rats by up-regulating the expression of microRNA-542-3p(miR-542-3p).Methods:The SD rats(8～10 weeks) successfully modeled with UC were divided into model group,mesalazine group,and Gegen Qinlian Decoction low-,medium-and high-dose groups,and the same batch of healthy rats with equal weight were used as blank control group,16 in each group.UC affected colon cells were digested and passaged into 24-well plates after entering the logarithmic growth phase,with 2×105 cells per well.The miR-NC,miR-542-3p,anti-miR-NC,and anti-miR-542-3p plasmids were transfected into colonic epithelial cells of UC rats by lip3000 and grouped.The miR-NC and miR-542-3p groups were treated with 10% normal saline for 24 h,while the anti-miR-NC and anti-miR-542-3p groups were treated with 10% Gegen Qinlian Decoction for 24 h.The changes of inflammatory mediators and oxidative stress-related indexes in the cell supernatants were analyzed after treatment.The contents of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6) and interleukin-8(IL-8) in rat colon tissue and cells were detected by enzyme-linked immunosorbent assay(ELISA).The contents of malondialdehyde(MDA),superoxide dismutase(SOD) and glutathione peroxidase(GSH-PX) were detected by chemical colorimetry.Real-time fluorescence quantitative PCR(RT-qPCR) was used to detect the expression level of miR-542-3p.Results:Compared with the conditions in model group,the contents of TNF-α,IL-6,IL-8 and MDA in Gegen Qinlian Decoction high-and medium-dose groups were decreased and the expression levels of SOD and GSH-PX and miR-542-3p were increased(all P<0.05).Compared with UC+miR-NC group,UC+miR-542-3p group had elevated expression level of miR-542-3p,SOD and GSH-PX and lowered contents of TNF-α,IL-6,IL-8 and MDA(P<0.05).Compared with the conditions in UC+anti-miR-NC+Gegen Qinlian Decoction group,the expression levels of miR-542-3p,SOD and GSH-PX in UC+anti-miR-542-3p+Gegen Qinlian Decoction group were reduced,and the contents of TNF-α,IL-6,IL-8 and MDA were increased(P<0.05).Conclusion:Gegen Qinlian Decoction protected UC rats by up-regulating the expression of miR-542-3p,thus eliminating intestinal inflammation. |
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